High flow, low hold up filters

ABSTRACT

Filter devices with a flow director surrounded by a filter element providing a large filter surface area and low fluid volumes for efficient filtration and high recovery of sample fluids. The filter device can be configured with dimensions and sealing surfaces to functionally interact as a pre-filter for common usage disposable filter flasks.

FIELD OF THE INVENTION

Filters and prefilters having a high surface area in a small volume.Filters made more efficient in flow through and recovery using a flowdirector inside a cylindrical filter to expose final small volumes to alarger filter surface area.

BACKGROUND OF THE INVENTION

It can be difficult to harvest conditioned media from a suspended cellculture product. In eukaryotic or prokaryotic cultures the total solidscan be quite high (e.g., 10%) and the required level of particulateclearance may be to sterilization levels (e.g., not more than 1 micron).The end phase culture can include aggregations of cells and debris thatquickly clog prior art filters. Previously, it has been nearlyimpossible to directly filter more than a small volume of cell cultureharvest before the end filter becomes clogged. Typically, researchershave first removed the bulk of cells and cell debris from a cell cultureby centrifugation before attempting end filtration. However, this isslow, expensive, and labor intensive. Even with a centrifugation step,filters often become clogged after only a small volume of filtrate haspassed through the filter. The researchers must then throw away thefirst filter and continue the operation with one or more additionalfilters, until the job is done.

One way to increase the amount of filtrate that can pass through a finalfilter and to delay clogging is to increase the area of the filtersurface. In Meyer et, al., (U.S. 5,232,595), filter surface is increasedby arranging a large amount of filter material in a pleated arrangementaround a central cavity. Sample fluid flows into the central cavity andis filtered radially, with the filtrate collected on the exterior of thefilter in a sealed cartridge. Still the problem of clogging remains.Moreover, there is a tendency for such filters to filter and clog moreat the bottom of the filter, particularly at the beginning and end offiltration. The pleated cylinder design presents significant recoveryefficiency issues, e.g., in harvesting the final sample volume (trappedat the clogged bottom of the filter, e.g., with the sample pump full ofair and no longer able to transfer liquids).

To partially mitigate these problems, most commercially availabledisposable filter units come with an optional “depth filter” of largerpore size to place in front of the final filter disk. This can help insome cases, but clogging is often only delayed by a small percentage ofadditional volume, and centrifugation is usually still required for mostuses. For example, see Davis et al, (US 2005/0191619) wherein aprefilter of greater pore size is placed in front of the final filter ofsmaller average pore size. Often, such filter systems clog rapidly withthe of bulk cells in the prefilter or with prefiltered sample stillclogging the final filter.

In view of the above, a need exists for filter systems that can filter alarger amount of sample before clogging. We believe it would bedesirable to have filter designs with smaller residual volumes, e.g., toincrease overall sample filtrate recovery. The present inventionprovides these and other features that will be apparent upon review ofthe following.

SUMMARY OF THE INVENTION

The filtration devices of the inventions provide, e.g., high volumefiltration flows, at lower pressure differentials, and with higheroverall recoveries. Substantial depth and/or surface areas of theprefilters can receive large amounts of crude samples while passing onrelatively clarified filtrates to a final filter for polishing withoutclogging. The present systems function so well in handling crude samples(such as end cell cultures or lysates) that a centrifugation step maynot be necessary preliminary to filtration.

A filtration device of the invention can include a filter (e.g., acylindrical, polygonal, or conical filter element) surrounding a filtercentral axis, a top end, a bottom end, an external filter surface, aninternal filter surface, and an internal space between the central axisand internal filter surface. Within the filter internal space, a flowdirector can be positioned to displace more of the internal space towardthe bottom end of the filter than it displaces toward the top of thefilter. At the end of a filtration, a central sleeve can be insertedinto the internal space to further displace retentate and improverecovery. Thus, the small volume of sample at the end of a filtrationprocess is exposed to a larger filter surface area than for conventionalfilters, e.g., for less clogging and higher recovery.

The filters can be of any type appropriate for the sample to befiltered. For example, the filter can be a filter sheet, a depth filter,a pleated sheet, an open cell foam, and/or the like. The filter pleatsor folds can be, e.g., directed horizontally or vertically. The filterscan be shaped, molded, pleated, folded gusseted, and or the like, tofunctionally fit into the filter device body, e.g., around the centralfilter axis. In many preferred embodiments, the filter can be made froma pleated sheet of filter membrane or fiber mat of filter material. Thefilter material is typically formed into a cylindrical, conical, orconic section shape (though it can have other appropriate shapes, e.g.,that laterally surrounds (e.g., encompass) the sample a fluid volume).In one embodiment, the filter is a pleated type with a generally conicalshape, e.g., splayed out toward the top of the filter. Alternately, thefilters can be splayed out toward the bottom. The filters are oftenemployed as prefilters (though they can be, e.g., the final or onlyfilter), e.g., with average pore sizes typically ranging from less thanabout 3 microns to more than 10 microns.

The flow directors can be any shape to direct sample inflows moreeffectively to the filter surfaces, as described herein. Flow directorsare typically not merely uniform tubes or internal housing supports ofcylindrical or conical filters. The flow directors are typicallyconfigured to provide a smaller remaining interior space (not occupiedby the director) between the flow director and filter at the bottom ofthe filter than the remaining interior space at or near the top of thefilter. For example, the flow director can have a parabolic ortriangular cross-section in a plane through the central axis of thefilter, with the narrower end up. Optionally, this feature (interiorspace decreasing in the direction of flow) can be obtained by the filterelement intruding more to the central interior space efferently (towarddistal flow or bottom) and/or the flow director taking up more of theinternal space efferently.

The filter devices often have a flange near the top of the filterelement to provide a structure for sealing and mounting of other deviceelements. For example, the device can include shoulder flange extendingradially out from the filter top end in a plane perpendicular to thecentral axis. Further, a sample reservoir can be mounted, e.g., to thetop side of the flange, and the bottom side can have a sealing surface(e.g., a resilient gasket). A bottom lateral flange can extend outradially from or near the bottom end of the flow director or housingbottom, with the flange comprising a top surface in sealing contact withthe filter bottom end. The flange can be configured to seal against thetop edge of a final filter reservoir. A spacing (ring) adaptor can bepositioned between the filter device and final filter, e.g., to allow alarger filter in the final filter reservoir. The flange can be planar,or, e.g., describe a conic section. Optionally, the filter housings canhave sealed side walls and connectors at the top and/or bottom(hermetically sealed system) to sealably attach to culture vessels,conduits, pumps, and/or receiving vessels.

A sample reservoir can be positioned above the filter top end andhermetically sealed to the filter top end or flange. The reservoirtypically has a volume ranging from less than about 10 ml to 10 L ormore or about 50 ml to 2 L. The filter can be sealed to the other filterdevice elements using, e.g., polymerizing resins, or hot meltthermoplastics.

The filter device can be positioned in a reservoir of a vacuum filterapparatus (such as a commonly available disposable final filter flask).A vacuum pump (optionally disposable) or filtrate pump can befunctionally integrated into the filter device or the vacuum filterapparatus. Optionally, the filter device can have a removable orintegrated manifold with a port configured for connection to a vacuumsource.

In an alternate aspect, the filter device can include a cylindrical orconical filter element mounted within a cylindrical or conical housingand comprising a central axis, a top end, a bottom end, an outersurface, an inner surface, and an inner cavity having a volume betweenthe central axis and filter inner surface. The housing can comprise acylindrical or conical body comprising a top, bottom, and sides; and, ahousing top end comprising a shoulder flange extending radially from acentral opening. The housing sides (e.g., below the flange) can beperforated to allow flow of filtrate out of the filter. The filterelement can, e.g., provide its own support, have a firm supportinglayer, or a support frame. The flange can extend from the centralopening across the filter top end, and extend radially in a plane beyondthe body. A housing bottom end can extends across the cylindrical bodybottom, sealing the filter exterior from the filter interior. Thehousing bottom end can include an outlet port for filtrate comingthrough an internal filter pack (e.g., positioned as a flow director).

The filter device can have a flow director comprising a top end and abottom end, and be mounted within the inner cavity. The flow directorcan have a smaller cross-section at the top end (e.g., end toward sampleinput) than at the bottom end (e.g., end toward filtrate outlet). Thatis, e.g., a vertical cross-section through the flow director can taperin from bottom to top. The flow director can have a verticalcross-section through a center describing a conic section. The radialdistance between the flow director top end and the filter can be, e.g.,at least two or three times a radial distance between the flow directorbottom end and the filter. The top end of the flow director can be,e.g., at least 0.5 inch below the filter top end, and the bottom end ofthe flow director not more than 0.25 inch from the filter bottom end.The flow director can be impermeable, or act as a second (internal)filter element.

The filter element can be a bulk filter, a depth filter, a membranefilter, and/or the like. In many embodiments, the filter is a pleatedsheet or quilt of filter membrane and/or fiber material. The filterelement can be adapted for filtration with an average pore size of 20microns, 5 microns, or less. The depth dimension (traversly through themedia) of the filter pleats or bulk filter media can range from lessthan about 0.05 inches to more than about 2 inches, or about0.1 inch toabout 1 inch. The diameter of the inner cavity can range from less thanabout 0.5 inches to more than 10 inches, or from about 1.5 inches toabout 3 inches. In some aspects, the filter element can include one ormore afferent filter layers inside an efferent filter element (e.g.,co-laminated), and the average pore size of the afferent filter layerscan be greater than an average pore size of the efferent filter element.Alternately, the filter element can have a pore size gradient ofincreasingly smaller pores from the inside to the outside of the filter.

The shoulder flange diameter can range from less than about 3 inches toabout 7 inches or more. The shoulder flange can extend radially from thehousing body a distance ranging from less than about 0.25 inches toabout 3 inches or more. The shoulder flange can also include a sealingsurface on a planar (or tapered) bottom side of the flange extensionbeyond the body. The flange bottom side can include a resilient sealmaterial (e.g., a gasket or silicone layer). The filter top end can befluidly sealed to the shoulder flange, while the filter bottom end isfluidly sealed to the housing bottom end. In some embodiments, thehousing bottom does not include an outer sealing surface, or the housingbottom does not include a flange extending radially beyond the housingbody.

The filter devices of the invention can be configured, e.g., to mountfunctionally within the receiver (sample reservoir) of a disposablefilter flask having an average pore size at least 50% less that anaverage pore size of the filter element. Optionally, the filter devicecan have a threaded connection or tubing fitting connection in sealedfluid contact with the afferent side of the final filter system. Suchsealed versions of the filters can be capsules and physically orientedany direction during use.

DEFINITIONS

Before describing the present invention in detail, it is to beunderstood that this invention is not limited to particular devices,which can, of course, vary. It is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting. As used in thisspecification and the appended claims, the singular forms “a”, “an” and“the” include plural referents unless the content clearly dictatesotherwise. Thus, for example, reference to “a surface” can include acombination of two or more surfaces; reference to “particles” includesmixtures of particles, and the like.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention pertains. Although any methods andmaterials similar or equivalent to those described herein can bepracticed without undue experimentation based on the present disclosure,preferred materials and methods are described herein. In describing andclaiming the present invention, the following terminology will be usedin accordance with the definitions set out below.

As used herein, the directional terms refer to common usage at locationson the surface of the earth. For example a top surface is above a bottomsurface. Horizontal is perpendicular to the force of gravity, andvertical is parallel to the force of gravity at a local environment.Typically filters of the invention filter liquid samples with the samplefluid top surface horizontal and the flow of filtrate generally downvertical and generally parallel to the central axis of the filter. Aninternal surface of a cylindrical or conical object is that surfacefacing the central axis of the object. Radial is a line originating atan axis and progressing outwardly. The radial of a cylinder isperpendicular to the axis of a cylinder. Although the filter devices aredescribed herein, e.g., with reference to a central filter axis being“vertical” this is a reference orientation for discussion of thedevices. We note that, particularly where the filter reservoir is notopen, the central axis of the filter can be other than “vertical”, asspilling can be eliminated, e.g., with capsular designs. We note thatthe axis of a filter element can be tilted to any angle, in use.

A “flow director”, as used herein is a body in the space between thecentral axis of a filter and the internal surface of the filter but notin contact with the filter along most of its surface or length insidethe filter. For example, where the filter is a hollow cylinder or hollowcone, the flow director would be located in the hollow interior space ofthe filter, e.g., mostly spaced away from the filter or mostly spacedaway from the inner folds of a pleated filter. The flow directorgenerally occupies a larger cross-section across the central axis at ornear the bottom of the filter than at or near the top of the filter. Inthis way, the flow director can function to wet a larger filter surfacewith a smaller amount of sample, e.g., as a filtration process reachesthe end. In some embodiments, the flow director can take up about thesame cross sectional space top to bottom internal to the filter element,but the interior space reduces efferently, e.g., because the filterelement intrudes more axially toward the bottom.

The “reservoir” of a filter system is the container that holds thesample on the afferent (proximal) side of the filter before it isfiltered. A sample reservoir can be a chamber above the top of thefilter element.

A “prefilter” is a filter in sequence before a final filter in afiltration process. Typically the prefilter has a larger average poresize than the final filter. Usually, the filtrate of the prefilter flowsdirectly onto the next filter of a process (e.g., final filter) withoutholding the prefiltrate as a process intermediate.

Two components of a device are “sealed” at their points of contact whenan associated working fluid does not pass through the points of contactunder working pressures, as is understood in the art.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic diagram of a filter device having a cylindricalfilter element.

FIG. 2 is a schematic diagram of a filter device having a conical filterand parabolic flow director.

FIG. 3 is an image of a filter device installed as a prefilter to adisposable filter flask unit.

FIG. 4 is a schematic diagram of exemplary 50 ml and 1 L filter systems.FIGS. 4A and 4B show perspective views of 50 ml and 1 L filters. FIG. 4Cshows a side view of a 1 L filter and FIG. 4D a top view of a 1 Lfilter. FIG. 4E shows a side view of a 50 ml filter and FIG. 4F a topview of a 50 ml filter.

FIG. 5 is a schematic diagram showing a cross-section of a Harvest 50filter system.

FIG. 6 is a schematic diagram showing a cross-section of a Harvest 100filter system.

FIG. 7 is a schematic diagram of an exemplary 10 L filter system.

FIG. 8 is a schematic diagram of a pleated supplemental filteroptionally placed between the filter device filter element and the finalfilter.

FIGS. 9A to 9D are exemplary diagrams of a center sleeve shaped todisplace retentate from the interior space of a prefilter, e.g., nearthe end of filtration, or for small samples. FIG. 9A is an outer view ofa representative center sleeve, e.g., adapted to reduce the internalfilter volume of many prefilters described herein. FIG. 9B is a sideview of the sleeve. FIGS. 9C and Dshow a top down view and axialcross-section of the sleeve.

FIGS. 10A to 10C are exemplary diagrams of a filter device having bothan inner and an outer filter element. FIG. 10A is a perspective view ofa prefilter having an outer filter element, and also an inner filterelement, e.g., in a position similar to that of a flow director in otherembodiments discussed herein. FIG. 10B shows a top down view of theprefilter. FIG. 10C shows an axial cross-section of the filter.

FIGS. 11A to 11C are exemplary diagrams of a course filter adapted to beplaced before the present prefilter devices to remove gross particles.FIG. 11A is a perspective view of a course filter having an outersealing ring surrounding a course laminated filter disk. FIG. 11B showsa top down view of the course filter. FIG. 10C shows an cross-section ofthe course filter.

DETAILED DESCRIPTION

Filter devices and methods are provided to efficiently filter liquidsamples having high levels of suspended solids with good recovery. Inmany cases, the filter devices can act as prefilters to clear enoughsolids from a cell culture so that a single finishing filter can fullyclarify and/or sterilize many-fold more product than without theprefilter.

Filter devices can include a conical or cylindrical pleated filterelement (or filter element of another shape surrounding a centralinterior) around a flow director to effectively direct liquid samplesfor high yields. Alternately the filter is not pleated (e.g., foam orflat) and/or the filter element has an alternate appropriate shape. Thefilter device can have flanges to seal against, e.g., a final filtersample reservoir, allowing the prefilter to use the same vacuum(pressure differential) source for prefiltration and final filtration ina single step. In some embodiments, the filter device can include a portto receive a vacuum source to draw filtrate into a container. Otherfilter devices include prefilter and/or post filter sealed threads offittings to provide sealed transfer of conditioned media from a culturevessel to a receiving vessel.

Methods include provision of the filter device, insertion of the deviceinto the sample reservoir of a final filter, filling the device withcell culture sample, and applying a vacuum to the final filter filtrate(efferent) side, whereby the bulk of cells and cell debris is capturedin the device without clogging, and the prefiltrate has so few solidsand particles that large volumes of conditioned media can be clarifiedor sterilized in the final filter. In most cases, a preliminarycentrifugation step is not required to obtain a high yield conditionedmedia harvest when using the filtration device. Alternately, filtratecan be forced through the filter device using vacuum, pumps, and/orgravity to urge fluids through the filters in systems sealed withconnectors (threads and pressure fittings) commonly available in thefield.

Filtration Devices. The efficient filtration devices can include acombination of complementary features that together simplify andincrease efficiency of filtration processes involving samples containinglarge amounts of particles. The devices can have, e.g., conical pleatedfilter elements in a housing providing structure and flanges for sealingto final filter reservoirs. The filters can include a flow director inthe interior space of the filter to direct sample uniformly onto thefilter and increase the filter surface area contact when sample volumesare reduced, e.g., at the end of a filtration step.

As shown in FIG. 1 , the filtration device 10 includes a housing 11supporting cylindrical pleated filter element 12. Inside the filterelement is a flow director 13 to direct fluid flows against the interiorsurfaces of the filter element. The housing includes flange 14 tosupport the filter device across the top of a disposable final filterreservoir. The bottom surface of the flange can seal against a finalfilter reservoir to take advantage of the same vacuum source as thefinal filter for both prefilter and final filter steps. The filterdevice can have a sample reservoir 15 to receive any desired volume ofsample for filtration.

I. The filter elements are typically configured with a generaltopography surrounding applied sample with internal filter surfaces. Forexample, the filter element will have an inflow port (e.g., top opening)to receive sample, however, once inside the filter element the sample issurrounded by 360 degrees of filter material. Conical or cylindricalfilter elements will have a circular cross section. Alternately, thefilters can surround the internal volume with other geometric shapes,e.g., having oval, triangular, square hexagonal, cross-sections, and/orthe like. So that the filter does not have to be sealed from theenvironment to prevent spillage, the inflow port is usually at the topof the filter element, allowing a technician to simply pour sample intothe filter element or into a reservoir above the inflow port.Alternately, the sample can be applied through a sealed fluid flowsystem.

The filter elements can surround sample, enhancing the area of filtersurface in a small volume. Filter elements can be shaped as a containeror as a conduit. The overall topography of the filter element can bedescribed, e.g., as a cylinder, cone, square stock conduit, cone withthe point end removed, a cup, a sphere, a parabolic trajectory, and/orthe like. In preferred embodiments, the filter element has a cylinder orconical shape; these shapes can provide enhanced surface to volumeratios and strength against pressure differentials. Where the filterelement has an open bottom end, the opening is typically sealed with ahousing floor and/or flow director bottom.

The filter element is often a depth filter capable of retaining orcapturing a large volume of particulate material. For example, thefilter material can be a deep layer of foam or randomly oriented fibers,with a relatively large average pore size. In some cases, the pore sizeof the material in the filter can become progressively smaller (gradientpore size) between the filter interior surface and exterior (efferent)surface. In a preferred embodiment, the filter is a pleated cylinder orcone. Pleated cones can be formed by splaying out the top end of apleated cylinder and sealing the open bottom end with a flow director orhousing bottom wall.

The present devices can act as primary or single (final) filters.Usually, the devices are configured to act as prefilters with depthfilter attributes or relatively course permeability to collect the macroand micro particles that would clog a final (e.g., sterilization)filter. When intended to act as prefilters, the filter elements willgenerally have an average pore size ranging from, e.g., more than 100 µmto less than 0.1 µm, more than 50 µm to less than 1 µm, from 30 µm to 3µm, from 20 µm to 5 µm, or about 8 µm. When intended as a final filterfor conditioned media the pore sizes will generally be smaller, e.g., inthe range from less than about 0.2 µm or less to 3 µm or more. In somecases, the filter element can have a pore size gradient, e.g., becomingsmaller from the filter interior surface to the exterior surface, e.g.,ranging from 100 µm to 5 µm across the thickness of the filter material.

Optionally, the filters themselves can have a course pre-filter, toremove cell and polymer aggregations. For example, the top opening ofthe filter element interior can include a course filter to keep largeaggregates from passing from the reservoir to the filter elementinterior space.

Although the filter elements can be long, they typically have height,width and depth at about the same order of magnitude. The height (e.g.,from top to bottom) can range from more than about 1 m to less thanabout 1 cm, from 50 cm to 2 cm, from 25 cm to 5 cm, or about 10 cm. Thewidth and depth are typically about the same for the filter elements,e.g., because of a general radial symmetry of laboratory filter systems.The width and or over all depth (vertical height in use) of the filterelement can range from more than about 60 cm to less than about 0.5 cm,30 cm to less than about 1 cm, from 25 cm to 3 cm, from 15 cm to 5 cm,or about 10 cm.

II. Flow directors fit inside the interior space of the filter elementand define an interior space for fluid flows between the director andthe filter element. The directors typically run from the area at or nearthe bottom of the filter element to an area at or near the top of thefilter element. Near the bottom of the filter element, the directorcomes closer, or contacts the filter element, leaving only a smallvolume for sample at the bottom of the filter. Nearer to the top of thefilter element the director typically occupies a smaller proportion (ornone) of the filter interior cross-section, thus leaving a larger volumeof space for sample near the top of the filter element.

Flow directors typically have a shape that gradually reduces in crosssection from the bottom to the top. For example, a flow director canhave a shape describing a pyramid, a cone pointing upward, a hemisphere,a parabolic dome, an inverted parabolic trajectory, and/or the like. Incertain preferred embodiments, the flow director is a parabolic domeextending from the bottom of the filter element to the level of thefilter element top. In other embodiments, the flow director does notchange cross sectional area from top to bottom, e.g., having acylindrical shape.

The flow directors generally take up less of the filter interior crosssection moving from bottom to top. That is, e.g., they are generallytapered. At the bottom of the filter element interior, the flow directormay contact the bottom of the filter element. At the top of the filterelement interior, the director may only occupy point space, or thedirector may not actually extend all the way to the top of the interiorspace. The bottom of the flow director can take up from about 100% toless than about 40% of the cross sectional area of the bottom of thefilter element, or from 99% to 50%, from 95% to 70%, from 90% to 80%. Inmost embodiments, the director takes up all or most of the crosssectional area of the interior space at the bottom of the filterelement. The top of the flow director can take up from about 0% to lessthan about 1% of the cross sectional area of the top of the filterelement, or about 2%, 5%, 10%, 25%, 50%, or more than 75% of the area.Flow directors typically take up more than half the volume inside thefilter element, or from more than 90% to less than 30%, from 80% to 40%,or about 75%. Preferred flow directors can take up more volume interiorto the filter element at the bottom half of the filter element than atthe top. In some embodiments, the flow director is uniform in crosssection but the internal space cross sectional area increases due toincreased spacing of the filter element away from the flow director.

The flow directors can have a height and width about the same, buttypically the directors are taller than wide. The height can range frommore than about 1 m to less than about 1 cm, from 50 cm to 2 cm, from 25cm to 5 cm, or about 8 cm. This can vary, e.g., according to the overallsize of the filter device involved. The presence of the flow directorcan allow filter elements to be longer while retaining good samplerecovery characteristics. The width and/or over all depth of the flowdirector can range from more than about 30 cm to less than about 1 cm,from 25 cm to 3 cm, from 10 cm to 4 cm, or about 5 cm.

The flow directors can be sealed against fluid flows at the bottom ofthe filter element. In one aspect, a flange extends out radially fromthe bottom of the director and the filter element is sealed to theflange and/or supported by the flow director flange, e.g., as shown inFIG. 2 .

III. The filter devices can include a housing (or frame members, e.g.,top and bottom), e.g., to provide structure and to integrate theelements of the device. For example, a housing can provide an overallstructure providing support to the filter element, which may not besturdy, particularly when wet. The housing can also have extending topand/or bottom flanges that can provide sealing surfaces. The housingscan provide structural support for mounting the flow director and/or asample reservoir.

In certain embodiments, the housing will have a flange extending out ata level at or near the top of the filter element area. This flange canact as a sealing surface for the top of the filter element, as amounting position for a sample reservoir, and/or to provide a contactsurface for sealing the device to the top of a final filter samplereservoir top edge. The flange can be planar. Alternately, the flangecan be tapered (e.g., up to about 30°), e.g., to aid in fitting andsealing to the reservoir of a final filter.

A bottom flange can extend out the bottom of the filter element area.This flange can act as a sealing surface for the bottom of the filterelement and/or as a mounting position the bottom of a flow director (orsecond central filter). In many cases, this bottom flange expands outand is mounted to the bottom of the flow director.

The housing can have structural elements unifying top and bottom deviceelements, and supporting the filter element. For example, a cylindricalor conical wall, screen, or cage can run between the top and bottomdevice elements. The structure can strengthen the entire device. Thestructure can prevent the filter element from bowing out under theforces of working pressure differentials. The structure can allowpermeation of filtrate from the filter into a collection vessel or intoa final filter sample reservoir. In certain embodiments, the “housing”includes top and bottom elements not connected with other housingstructures (walls, rods, etc.) but are connected through the filterelement structures.

In certain embodiments, the housing side wall is not perforated butprovides a sealed space between the wall and exterior surface of thefilter element. The housing can include threaded fillings or sealedconnectors providing a sealed filtrate flow path, e.g., through tubing,additional filters, fluid pumps, and/or filtrate receiving vessels.

IV. Reservoirs can be provided to retain additional volumes of samplewhile filtration is in progress. Optionally, the filter element interiorspace can act as a reservoir for sample to be filtered. As discussedabove, the sample reservoir can be mounted, e.g., to the top of thefilter element or a top housing flange to provide additional sample asearlier sample filtrate flows through the filter element. The fluids inreservoirs are typically intended to be gravity fed to the filter.

Depending on the size of the filter element, and expected clogging rate,the reservoir can vary considerably in size. In many instances, thereservoir has a volume approximately the same as a final filterreservoir or as the final filter filtrate collection vessel with whichthe filter device is paired as prefilter.

In some embodiments, there is no substantial reservoir space above thefilter element, e.g., with sample being pumped (or gravity fed) to thefilter element interior directly from the culture vessel, or through atransfer conduit.

V. Additional aspects. The filter device can be paired with a finalfilter to act as a prefilter. The most common final filters are theubiquitous commercially available disposable flask filters. Thesefilters include a disk filter between an upper sample reservoir and alower filtrate collection flask. The filter is in a housing thatincludes a side arm conduit providing a path for a vacuum source toevacuate the collection flask, creating a pressure differential to drivethe filtration process. For example, see filter flasks by NALGENE™ andMILLIPORE™. These disposable filter units range in capacity from about50 ml to 100 ml, 150 ml, 250 ml, 500 ml, 1 L, 2 L, or more. One thingthese disposable filter units have in common is an upper reservoir(“funnel”) with a top edge in a horizontal plane. The upper flanges ofthe filter devices described herein can rest and seal against thedisposable filter top edge so that filtration by the filtration deviceprefilter can be accelerated by the same vacuum used to drive filtrationin the disposable final filter.

In another aspect, the present filter or the disposable final filter canbe provided with a dedicated vacuum or pressure pump. That is, insteadof providing vacuum from a “house vacuum” system through a vacuum hose,the present filter system can be powered by an attachable, ordisposable, vacuum pump. In a preferred embodiment, the vacuum pump isconfigured to provide a greater pressure differential than a typicalhouse vacuum system. The disposable pump can comprise a simpledisposable working pump head (e.g., diaphragm, vane, piston) adapted touse with a non-disposable power unit.

In an alternate configuration of the filter device, filtration is bygravity flow, without the need to provide an additional pressuredifferential across the filter element.

The filter devices described herein could act as final filters, e.g.,incorporated as the filter element in a disposable filter unit. That is,the present filter could replace the disc filter commonly provided incommercial disposable filter systems. The housing act as a manifold andinclude a side arm conduit allowing evacuation of air from a bottomfiltrate vessel.

Although the present discussions refer mostly to cell culture samples,the present devices and methods are not so limited. The filter devicesare suitable for filtration of any fluid containing any particles. Forexample, the filter devices can be used to filter particles out of air,to filter lubricants and fuels for machinery, to filter fluids in foodprocessing methods, and/or filter process intermediates or finalproducts in any number of industrial processes.

It is envisioned that an alternate filtration device can be configuredwith the flow director and filter element exchanging places. Forexample, in FIG. 1 , the flow director of 13 could be exchanged with asimilarly located filter and the filter 12 could be a flow director wallthat provides, e.g., the continuously reducing sample space from the topof the filter to the bottom.

VI. Filtration Methods. The filtration devices described herein can beused in a variety of ways. They can be primary standalone filters, e.g.,run by gravity or by a pressure differential. Because of the largefilter surface area and efficient fluid flow direction, the devices canalso act as prefilters receiving difficult samples, e.g., containinglarge amounts of particles or cells.

In one aspect of the methods, the bottom surface of a housing top flangecan be placed over the mouth of a vacuum vessel, sample poured into thefilter interior space, and a vacuum applied to the vessel to collectfiltrate into the vessel. Alternately, the filtration device canincorporate a fitting, such as a threaded fitting to screw-fit onto thevacuum vessel, or to a filtrate pumping system.

Optionally, the filter devices can be used as prefilters. Expressionhost cells can be grown to near end of growth phase in liquid suspensionculture. A filtration device described herein can be placed so that thefilter element is in the sample reservoir of a final filter system, withthe top flange sealing against the top edge of the final filterreservoir. The suspension culture (e.g., without previous centrifugationto pellet cells) can be poured into the sample reservoir of the filterdevice and a vacuum applied to the filtrate collection vessel of thefinal filter system. The lower pressure in the collection vessel candraw air out of the space between the final filter dick and the presentfilter device, forcing a tighter seal between the top flange and finalfilter top edge. As a relative vacuum develops in the space, conditionedmedia is drawn through the filter device filter element to collect inthe final filter sample reservoir. Because most of the cells are removedfrom the conditioned media, the entire filtrate collection vessel can befilled through the final filter disk without a clogging incident.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Example 1 - Conical Filter With Parabolic Flow Director

A pleated filter is splayed into a generally conical shape thatcooperates with a flow director to provide the tapered sample spacebetween the filter and flow director, as shown in FIG. 2 . The designfurther provides more external space at the bottom where greaterfiltrate flows are expected to be experienced. In addition, the designenhances initial flows nearer the top of the filter element so that lessmaterial accumulates at the bottom of the filter, thus aiding infiltration and collection of residual sample volumes at the end of afiltration step.

A filtration device 20 includes a conical pleated filter element 22.Inside the filter element is a flow director 23 to direct fluid flowsagainst the interior surfaces of the filter element. The housingincludes upper flange 24 providing a seal 26 to interact with the topedge of an appropriate disposable filter flask, to seal the top of thefilter element, and on which to mount sample reservoir 25. The housingcan further include ribs (not shown) running within some filter pleatsas reinforcement against pressure differentials experienced duringfiltration processes.

Example 2 - Filter Device Pre-Filtering for a Disposable Final Filter

The filter devices described herein can be inserted into and into afinal filtration flask system to act as a prefilter.

As shown in FIG. 3 , a filter device 30 described herein was placed intoa disposable final filter system 31 sample reservoir 32 with upperflange 33 in sealing contact with the top edge of the final filtersample reservoir.

Example 3 - Exemplary Culture Harvest Filter Devices

Harvest Max 50 and Harvest Max 100 filter systems are presented in FIGS.4A to 4F. Scale is in [inches] and milimeters.

The Harvest Max 50 filter 40 is sized to receive 50 ml of sample intosample reservoir 41. The inside surface of the reservoir can includeinternal threads adapted to functionally interact with the male externalthreads of a culture bottle, e.g., for direct sealed transfer of cellculture sample form a culture vessel to the filter input (afferent)side.

In use, a cell culture is completed in a culture flask, and thereservoir end of the filter device is attached and sealed to the topopening of the culture flask. Next, a vacuum flask is sealed to theflange around the bottom of the filter reservoir. The assembly is stoodwith the culture flask inverted on top of the filter and vacuum flask.Sample flows into the filter reservoir and space between the filterelement and flow director. Sample filtrate flows through the filter andinto the receiving flask at the bottom of the assembly.

Alternately, the filter device can have a non-perforated housingentirely around the exterior of the filter element 42. Further, thebottom 43 of the housing can be sealed from the interior of the filterelement but open to the space between the exterior of the filter elementand the non-perforated housing wall. The housing bottom can also includea threaded surface or seal adapted to sealably attach to a receivingvessel, such as a vacuum flask. Flow from the culture flask, through thefilter, and into the receiving vessel can be by force of gravity,relative vacuum (less than atmospheric pressure) in the receivingvessel, and/or a pressure pump on the afferent side of the filter.

A cross section of a Harvest Max 50 filter is presented in FIG. 5 .Note, an optional multi-layer pre-filter insert 44 can be mounted overthe filter interior space, e.g., to remove agglomerations of cells. Theprefilter itself can be pleated, as shown in FIG. 8 . Pleats of the mainfilter element 45 are finer depth filter media with pores scaled toremove the bulk of remaining cells, cellular debris, and finerparticles.

The Harvest Max 100 filter 50 is sized to receive up to 1 L of sampleinto sample reservoir 51. The inside surface of the reservoir caninclude internal threads adapted to functionally interact with the maleexternal threads of a culture bottle, e.g., to directly receive samplefrom an attached culture bottle or pump hose adaptor. The Harvest Max100 can be configured to function as per the Harvest Max 50, describedabove.

A cross section of a Harvest Max 50 filter is presented in FIG. 6 .Note, an optional multi-layer pre-filter insert 52 can be mounted overthe filter interior space, e.g., to remove agglomerations of cells.Pleats of the main filter element 53 are finer depth filter media withpores scaled to remove the bulk of cells, cellular debris, and finerparticles. The outlet at the bottom of the filter housing can have,e.g., internal threads to screw on to a filtrate flask. Alternately, thefilter bottom can seal to the top of a final filter. Optionally thebottom can have fittings to interact with a pump, e.g., with hoses totransfer filtrate to a desired receiving vessel.

Note the filter device is resting in the reservoir 55 of a final filterunit having final filter 56.

Example 4 - Exemplary Filter Devices

The Harvest Max 1000 filter system is presented in FIG. 7 .

The system includes the filtration unit 70 and optional single use pumphead 71. The Harvest Max 1000 can receive a liter into reservoir 72, andcan also have samples pumped into the reservoir through input port 73.The filter device typically has the filter capacity to filter up to,e.g., 10 liters of uncentrifuged end phase eukaryotic cell culture.

In use, sample can be applied to the sample reservoir to flow by gravityinto a receiving vessel. Optionally, the sample can be drawn through thefilter by a relative vacuum in a receiving vessel, or driven by theforce of a pump installed between the filter outlet and receiving vesselinlet.

Note the filter device can be configured to have a housing (skirt)coming down from the reservoir to surround a top portion of the filterelement, while the final filter reservoir surrounds a bottom portion ofthe filter element. In this way, a longer filter element can be providedthan would fit within the height of the final filter reservoir.Optionally, filter devices having filter elements too long to fit in thereservoir can be placed upon an adaptor ring or cylinder that sealablyspaces the filter device an appropriate distance above the final filterreservoir.

In many embodiments, e.g., wherein the culture vessel is large orungainly (e.g., 10 L culture bags), a pump can be used to continuouslyor intermittently fill the filter sample reservoir. Such a pre-reservoirpump can optionally provide the force driving filtrate through thefilter element. Optionally, sample can be received by gravity fromculture containers mounted above the filter device.

Example 5 - Supplemental Filter Before the Final Filter

Supplemental filters can be located between the filter element and finalfilter. The supplemental filter is typically a horizontal diskconfigured to mount or rest above final filter.

FIG. 8 shows an exemplary supplemental filter 80 for mounting across thefiltrate flow below the filter device filter element. The filtermaterial 81 is pleated and surrounded with a supportive collar 82. Thissupplemental filter can be used to further assist the HarvestMax toincrease throughput. The average pore size of the supplemental filter istypically between the average pore size of the filter device filterelement and the average pore size of the final filter. The supplementalfilter can have any appropriate average pore size, e.g., 0.2 µ or 0.45 µor 0.65 µ membrane, pleated to have ~100 cm² area or more to furtherprotect the final filter in the flask with limited area i.e. 60 cm2.

Example 6 - Attachments to Standard Culture Vessels

Cell culture products, such as 5L shaker bottle containers, can havetransfer caps for direct connection to the afferent side of the filterdevices, or connection through tubular connections. One or two of their5L units can be harvested one at a time with a connector of ⅜″ Legrisfitting to ⅜″ clear vinyl or other tubing with a ⅜″ Female QDC quickdisconnect connector from Harvest Max 1000 to the top of the 5 L whichhas a ⅜″ MQDC. Typically, a direct connect for gravity harvest toclarification can be accomplished.

Also with the Harvest Max 1000 it is important to note a direct harvestcan be made from a G.E. or other 22 L (10 L working volume) cell culturebag via a female ⅜″ CPC connector on the bag and an adaptor from the topof the Harvest Max (Legris × ⅜″ MQCD).

Example 7 - Center Sleeve

At the end of a filtration, even with the low retentate filtersdescribed above, there can come a point where the level of sampleretentate drops below the top of the filter element. Flow rates canslow, air may bubble through the uncovered top filter surfaces, andremaining retentate may contact only the lower filter element, wherepores may be clogged with debris.

Filters with flow directors, as described above, have low dead volumeson the sample side of the filter, compared to standard filter systems.However, for particularly small or precious samples, technicians may betempted to rinse the retentate side of the filter to chase outadditional sample. However, this will dilute the filtrate.

To solve this problem, we have developed a center sleeve device to takeup the internal volume, e.g., between the filter element and flowdirector element. As shown, e.g., in FIG. 9 , a center sleeve 90 can beprovided with a shape closely approximating the internal space betweenthe filter and flow director.

The center sleeve can be any shape that effectively displaces asignificant proportion of retentate from the interior volume of thefilter section of the filter device. For example, the sleeve internalsurface 91 can be shaped to uniformly contact the flow director surface,e.g., leaving no volume between the flow director and sleeve. The sleevecan be shaped to so that the outer sleeve surface 92 uniformly contactsthe filter element internal surface, or to contact inner folds of thepleated filter, e.g., leaving a minimum volume of retentate between thepleats.

Alternately, depending, e.g., on the amount of debris or flowcharacteristics of the sample, the sleeve can be shaped to leave a smallspace between the sleeve and filter. For example, an installed sleevecan leave between less than 0.1 mm and 10 mm or more of clearancebetween the outer sleeve and filter element inner surface; between 0.3mm and 5 mm, between 0.5 mm and 3 mm, or about 1 mm.

In use, a prefilter device can be placed into the reservoir of a finalfilter, and sample poured into the sample reservoir of the prefilterdevice. With vacuum applied to the final filter, sample is drawn pastthe flow director and through the prefilter filter element. As thesample retentate is reduced in volume, the top meniscus will drop belowthe top of the filter element. Filtration will slow due to the reducedtotal filter surface contact, and tendency of the lower filter to bemore clogged than the upper filter element. At the filtrationtechnician’s discretion, the center sleeve can be inserted into theinterior space of the prefilter device, e.g., between the flow directorand filter element, displacing retentate up to higher levels of thefilter element. Filtrate flow will increase, entry of air into filterpores is avoided, and the filtrate harvest efficiency will be increased.

Example 8 - Filters With Inner and Outer Filter Elements

The amount of filter surface area in the prefilter devices can beincreased by, e.g., adding a filter pack to the space interior to thefirst outer filter element. For example, the flow director can be afilter element.

As shown in FIG. 10 , the prefilter device 100 can include an innerfilter 101 inside of the outer filter element 102. Sample introduced toreservoir 103 to pass between the inner and outer filter elements intoinner space 104. Filtrates can pass through either the inner or outerfilter element. Filtrate passing through the inner filter element 101can pass, e.g., into the reservoir of a final filter through filtrateexit port 105. Note, the inner volume 104 between the inner and outerfilter elements can be tapered toward the bottom of the filter device ina manner similar to filters described herein having a flow director.

Example 9 - Adaptor to Allow Prefilter Use on Smaller Final Filters

It can be desirable to prefilter a small volume into a smaller finalfilter, e.g., having a smaller sample reservoir. The problem can be thatthe prefilter element may be too long to allow the prefilter seal flangeto contact the final filter reservoir opening.

In such a case, an adaptor ring can be used to fill the space betweenthe prefilter sealing flange and the top of the final filter reservoir.Such a ring can include resilient sealing surfaces on the top and/orbottom surface to aid in sealing at the adaptor prefilter interface andthe adaptor final filter reservoir interface.

Example 10 - Course Filter Before Prefilter

In certain cases, e.g., where input suspensions include significantamounts of clumped or aggregated materials, flows through the prefilterdevice can be improved by including a course filter disk afferent to theprefilter. In many cases a course disk 111 (see FIG. 11 ) can bepositioned, e.g., across the bottom of the prefilter reservoir (entry tofilter element internal space) to remove gross particles before they cancontact the prefilter filter element.

In an embodiment, a course filter ring 111 included a triple laminatedcotton filter disk 112 mounted central to a sealing ring 113. The filterdisk should have a larger average pore size than the distal prefilterelement. The ring can include a resilient sealing surface.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims.

While the foregoing invention has been described in some detail forpurposes of clarity and understanding, it will be clear to one skilledin the art from a reading of this disclosure that various changes inform and detail can be made without departing from the true scope of theinvention. For example, all the techniques and apparatus described abovecan be used in various combinations. All publications, patents, patentapplications, and/or other documents cited in this application areincorporated by reference in their entirety for all purposes to the sameextent as if each individual publication, patent, patent application,and/or other document were individually indicated to be incorporated byreference for all purposes.

What is claimed is:
 1. A filter device comprising: a filter elementsurrounding a central axis and comprising a top end, a bottom end, anexternal filter surface, an internal filter surface, and an internalspace between the central axis and internal surface; a flow directorpositioned within the internal space and displacing more of the internalspace toward the bottom end of the filter than the flow directordisplaces toward the top of the filter; and, a reservoir hermeticallysealed to the filter top end through a shoulder flange extendingradially out from the filter top end in a plane substantiallyperpendicular to the central axis.
 2. The device of claim 1, wherein thefilter is a pleated filter.
 3. The device of claim 1, wherein thereservoir has a volume greater than a volume of the internal space. 4.(canceled)
 5. The device of claim 1, wherein the flow director has aparabolic cross-section in a plane through the central axis. 6-9.(canceled)
 10. The device of claim 1, wherein a bottom end of the flowdirector comprises a lateral flange extending out radially, which flangecomprises a top surface in sealing contact with the filter bottom end.11. (canceled)
 12. The device of claim 1, positioned in a reservoir of avacuum filter apparatus.
 13. The device of claim 1, further comprising acentral sleeve shaped to fit into an interior space between the filterelement and the flow director, whereby positioning the sleeve into theinterior space displaces sample up to provide improved contact with thefilter element. 14-21. (canceled)
 22. The filter of claim 1, wherein theshoulder flange further comprises a sealing surface on a planar bottomside of the flange extension beyond the body. 23-28. (canceled)
 29. Thefilter of claim 1, wherein a radial distance between the flow directortop end and the filter is at least two times a radial distance betweenthe flow director bottom end and the filter. 30-32. (canceled)
 33. Thefilter of claim 1, further comprising a prefilter (depth filter) elementextending across the filter device reservoir positioned to prefiltersample before it contacts the first filter element.
 34. The filter ofclaim 1, wherein the device is configured so that a sample liquidapplied to the flow director top end wets the filter top end.
 35. Thefilter of claim 1, wherein the flow director contacts the filter elementat most only at the filter bottom end.